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More than twenty years ago, scientists succeeded in pushing the limits ofoptical detection to single molecules using fluorescence. This breakthrough hasrevolutionized biophysical measurements, but restrictions in photophysics andlabeling protocols have motivated many efforts to achieve fluorescence-freesingle-molecule sensitivity in biological studies. Although several interestingmechanisms using vibrational spectroscopy, photothermal detection, plasmonicsor microcavities have been proposed for biosensing at the single-protein level,no method has succeeded in direct label-free detection of single proteins.Here, we present the first results using interferometric detection ofscattering (iSCAT) from single proteins without the need for any label, opticalnanostructure or microcavity. Furthermore, we demonstrate super-resolutionimaging of protein binding with nanometer localization precision. The ease ofiSCAT instrumentation promises a breakthrough for industrial usage as well asfundamental laboratory experiments.

Direct optical sensing of single unlabeled small proteins and super-resolution microscopy of their binding sites